Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia; Bioavailability Laboratory, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia.
Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia; Bioavailability Laboratory, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia.
J Pharm Biomed Anal. 2019 Feb 5;164:1-8. doi: 10.1016/j.jpba.2018.10.005. Epub 2018 Oct 4.
Suvorexant is a novel sedative/hypnotic drug approved for treatment of insomnia. It has significant forensic importance due to its hypnotic and depressant effects on central nervous system. In this study, a highly sensitive UPLC-MS/MS assay was developed and validated for the determination of suvorexant in urine sample. A simplified dispersive liquid-liquid microextraction followed by ultrasound assisted back extraction from solidified floating organic droplets was employed for sample preparation. The 20 μL of 1-undecanol and 200 μL of acetonitrile were used as extraction solvent and dispersive solvent, respectively. An ultrasound assisted back extraction step was employed to enable the cleanup procedure compatible with mass spectrometric detection. Acquity CSHC column with mobile phase composition of 15 mM ammonium acetate: acetonitrile: formic acid (15:85:0.1%; v/v/v) were used for chromatographic separation. The multiple reaction monitoring transition of 451.12 →104.01 and 451.12→186.04 were used for identification and quantification of suvorexant, respectively, whereas 237.06→194.1 was used for IS in positive mode. The assay demonstrated good linearity in the range of 0.27-1000 ng mL with limit of detection (LOD) and quantification (LOQ) of 0.10 and 0.27 ng mL, respectively. Assay validation was performed by following SWGTOX guidelines and all validation results were found to be within acceptable limits. This is the first report of dispersive liquid-liquid microextraction based on solidification of floating organic droplets employed to UPLC-MS/MS for application in biological fluids.
苏沃雷生是一种新型镇静/催眠药物，用于治疗失眠症。由于其对中枢神经系统的催眠和抑制作用，它具有重要的法医学意义。在本研究中，开发并验证了一种用于尿液样品中苏沃雷生测定的超高效液相色谱-串联质谱法。采用简化的分散液-液微萃取，随后通过超声辅助从固化的浮质有机液滴中进行反向萃取，用于样品制备。20 μL 的十一烷醇和 200 μL 的乙腈分别用作萃取溶剂和分散溶剂。采用超声辅助反向萃取步骤，使净化程序与质谱检测兼容。采用 Acquity CSHC 柱，流动相组成为 15 mM 乙酸铵：乙腈：甲酸（15:85:0.1%；v/v/v），用于色谱分离。451.12→104.01 和 451.12→186.04 的多重反应监测转换分别用于苏沃雷生的鉴定和定量，而 237.06→194.1 则用于正模式下的 IS。该测定法在 0.27-1000ng mL 范围内具有良好的线性关系，检测限（LOD）和定量限（LOQ）分别为 0.10 和 0.27ng mL。按照 SWGTOX 指南进行了测定验证，所有验证结果均在可接受范围内。这是首次报道基于固化浮质有机液滴的分散液-液微萃取用于 UPLC-MS/MS 在生物流体中的应用。
